Coated plates for fluorescence imaging of baculovirus-infected insect cells?

Mike Autry via methods%40net.bio.net (by jma from ddt.biochem.umn.edu)
Tue Apr 1 18:37:06 EST 2008


Methods:

Which coating is best to keep baculovirus-infected insect cells adhered to microscope plates: poly-lysine, integrin, laminin, etc? Any recommendations on plate type and vendor availability?

I have a major problem doing FRET microscopy on Sf21 insect cells infected with recombinant AcNPV baculoviruses:  following infection, the Sf21 cells detach from the glass microscope plate and roll around, especially when I change filter cubes (donor, acceptor, FRET dichroics) during photobleaching experiments.

I am expressing ER membrane proteins tagged with cyan and yellow fluorescent proteins (CFP, YFP).

Thanks,

J. Michael Autry, PhD
Research Associate, University of Minnesota
Minnesota Muscle Laboratory, David D. Thomas, PI
http://ddt.biochem.umn.edu/



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